|Anti-HGF Stable Cell Line
- Cell Line Description
- The protein encoded by this gene is produced by macrophages and has antiviral activity. This gene is intronless and the encoded protein is secreted. IL-22 is an α-helical cytokine. IL-22 binds to a heterodimeric cell surface receptor composed of IL-10R2 and IL-22R1 subunits. IL-22R is expressed on tissue cells, and it is absent on immune cells.
- Growth Properties
- Complete growth medium: Serum-free Medium
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
- Starting Cells From Frozen Cell Stock
- 1. Remove the packaging cell lines from liquid nitrogen and carry out a quick thaw. Float the cells in the 37°C water bath for 2 minutes until nearly (80%) thawed.
Once cells are thawed, it is important to dilute the cells 1:10 in growth media immediately to reduce the potentially toxic effects of the DMSO preservative on the cells.
2. Clean the outside of the vial with 70% ethanol before opening.
3. Remove the cells from the vial and add slowly into a 15ml conical tube containing 10 ml pre-warmed media.
4. Centrifuge for 3 minutes 1000 xg to pellet cells and remove the supernatant.
5. Add 14 ml of media and transfer cells to a T25 flask or a 100 mm culture dish.
6. Place the cells in the 37°C incubator with 5% CO2.
7. Allow incubation for 3-4 days to reach confluence. The cells will re-attach to the surface over a period of several days in culture at 37°C.
- 1. Centrifuge for 5 minutes 300 xg and discard culture medium.
2. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
3. Add appropriate aliquots of the cell suspension to new culture vessels.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
- Mycoplasma Status: Negative (MycoAlert Kit)
- Freeze Medium
- Complete growth medium 90%; DMSO, 10%
- Safety Considerations
- The following safety precautions should be observed.
1. Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
2. No eating, drinking or smoking while handling the stable line.
3. Wash hands after handling the stable line and before leaving the lab.
4. Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.
5. All waste should be considered hazardous.
6. Dispose of all liquid waste after each experiment and treat with bleach.
- Product Purity
- HPLC > 98%, One strip for SDS-PAGE.
- Humanized (from mouse) IgG1 - kappa
- Product Molecular Mass
- Product Storage
- It should be stored in liquid or vapor phase nitrogen. Reconstituted antibody aliquots should avoid repeated freeze-thaw cycles.
- Involvement in Disease
- Ficlatuzumab (formerly known as AV-299) is a potent hepatocyte growth factor (HGF) inhibitory antibody that binds to the HGF ligand with high affinity and specificity to inhibit HGF/c-Met biological activities. It is now in Phase II study for non-small cell lung cancer.
- Ficlatuzumab is a humanized monoclonal antibody designed for the treatment of cancers.
- HGF; hepatocyte growth factor (hepapoietin A; scatter factor); deafness, autosomal recessive 39 , DFNB39; hepatocyte growth factor; F TCF; fibroblast derived tumor cytotoxic factor; hepatopoietin A; HGFB; HPTA; lung fibroblast derived mitogen; scatter factor; SF; hepatopoeitin-A; hepatopoietin-A; lung fibroblast-derived mitogen; fibroblast-derived tumor cytotoxic factor; F-TCF; DFNB39;
- Chromosome Location
- Arf6 signaling events, organism-specific biosystem; Cytokine Signaling in Immune system, organism-specific biosystem; Cytokine-cytokine receptor interaction, organism-specific biosystem; Cytokine-cytokine receptor interaction, conserved biosystem; Direct p53 effectors, organism-specific biosystem; FGF signaling pathway, organism-specific biosystem; Focal Adhesion, organism-specific biosystem;
- catalytic activity; growth factor activity; protein binding; protein heterodimerization activity; NOT serine-type endopeptidase activity;
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